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Mitochondrial oxidative phosphorylation became functional under aglycemic hypoxia conditions in A549 cells

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Abstract

Background

Normal cells produce energy (ATP) through mitochondrial oxidative phosphorylation in the presence of oxygen. However, many of the cancer cells produce energy with accelerated glycolysis and perform lactic acid production even under normoxic conditions called “The Warburg Effect”. In this study, human lung carcinoma cells (A549) were incubated in either a normoxic or hypoxic environment containing 5 mM glucose (Glc 5), 25 mM glucose (Glc 25), or 10 mM galactose (OXPHOS/aglycemic), and then the bioenergetic pathway was anaylsed.

Methods and results

HIF-1α stabilization of A549 cells with different metabolic conditions in normoxia and hypoxia (1% O2) was determined using the western blot method. After that, l-lactic acid analysis, p-PDH/PDH expression ratio, ATP analysis, and citrate synthase activity experiments were also performed. It was determined that HIF-1α stabilization reached the maximum level at the 4 h. It has been found that glycolytic cells produce approximately five times more lactate than OXPHOS cells under both normoxia and hypoxia conditions and also have a higher p-PDH/PDH ratio. It has been determined that citrate synthase activity in hypoxia of all metabolic conditions is lower than normoxia. It has been determined that Glc 5 and Glc 25 cells have more ATP production under normoxia than Glc 5 and Glc 25 cells in hypoxia. OXPHOS cells have showed more ATP production in hypoxia.

Conclusion

It has been determined that oxidative phosphorylation became functional in a hypoxic aglycemic environment despite the metabolic programming regulated by HIF-1α. This data is important in determining targets for therapeutic intervention.

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Data availability

The data that support this study are available from the corresponding author upon reasonable request.

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Funding

This work was funded by a grant from the Anadolu University (Project Nos. 1809S298, 1905S059).

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Pharmacy, Anadolu University, Eskisehir, Turkey

    Yüksel Öğünç Keçeci & Zerrin İncesu

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  1. Yüksel Öğünç Keçeci
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  2. Zerrin İncesu
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Contributions

YÖK—Data analyzing and draft manuscript preperation, ZS—Supervision of the research and critical revison of the paper, YÖK and ZS—Final approval of the version to be published.

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Correspondence to Yüksel Öğünç Keçeci.

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Öğünç Keçeci, Y., İncesu, Z. Mitochondrial oxidative phosphorylation became functional under aglycemic hypoxia conditions in A549 cells. Mol Biol Rep 49, 8219–8228 (2022). https://doi.org/10.1007/s11033-022-07400-6

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  • DOI: https://doi.org/10.1007/s11033-022-07400-6

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